Topical composition enhancing healing of viral lesions

ABSTRACT

Multiple daily applications of a topical composition having as the active ingredients an anesthetic and a surfactant with anti-viral activity decrease the time of healing of herpes simplex viral lesions from approximately seven to ten days to five to six days, as well as decrease inflammation and pain. The composition is applied to the lesions daily, approximately every four hours, preferably beginning at the prodromal stage of infection. Relief is almost immediate, and is characterized by decreased pain, swelling, and more rapid healing as compared with the untreated lesion.

This application is a continuation of application Ser. No. 07/985,279,filed Dec. 4, 1992, (U.S. Pat. No. 5,380,754 which is acontinuation-in-part of U.S. Ser. No. 07/887,101 filed May 19, 1992(abandoned), which is a continuation of 07/718,005 filed Jun. 20, 1991(abandoned), which is a continuation-in-part of Ser. No. 07/504,683filed Apr. 4, 1990 (abandoned), which is a continuation of Ser. No.07/161,802 filed Feb. 29, 1988 (abandoned).

BACKGROUND OF THE INVENTION

Herpes simplex virus (HSV) causes infections characterized byinflammation of skin and mucosal membranes of the mouth and genitalareas. The type of HSV most commonly associated with lesions of themouth and facial region is Type 1 HSV. Type 2 HSV is most commonlyassociated with lesions of the genital region.

The symptoms of HSV generally include erythema, inflammation, pain, andlesions. Symptoms initially include tingling, pain, and/or swelling atthe site of infection, followed by lesion formation (papules andvesicles) with scar formation and healing occuring in approximately tento fourteen days. After the initial infection, recurrent attacks mayoccur throughout life in response to various stimuli, including changesin body temperature, stress, ultraviolet radiation and exposure tochemicals.

Treatments for HSV infections are limited in efficacy. There is no knowncure at this time, although various anti-viral compositions are beingmarketed with claims of beneficial effects. Most anti-viral compoundscurrently in use are synthetic nucleoside analogs which interfere withthe replication of viral DNA or RNA. The mode of action of thesecompounds is to directly interfere with the action of DNA or RNApolymerase or chain termination after incorporation into specificnucleic acid polymers.

Many synthetic nucleoside analogs have been tested for the ability toinhibit viral replication. However, most of these have unacceptabletoxicity and cannot be used for treating humans. Interestingly, manyuseful nucleoside analogs have principal activity against the herpesgroup of viruses which include varicella-zoster virus, herpes simplexvirus, Epstein Barr virus and cytomegalovirus.

Currently, there is only one commercially available FDA approvednucleoside analog which has been shown to be efficacious in thetreatment of lesions of the skin produced by herpes simplex virus. Thiscompound is produced by Burroughs Wellcome Company and has been giventhe name Acyclovir. Acyclovir has been shown to be effective in thetreatment of primary genital herpes and, in many individuals, can reduceor prevent prevent recurrences when taken on a daily basis over longperiods of time. Acyclovir (acycloguanosine) is administeredsystemically for treatment of HSV with some success. Topical acyclovirhas been found to be somewhat effective for the primary lesion but notfor recurrent lesions. There is a wealth of clinical and scientificliterature relating to the use of acyclovir in herpes infection.

Interferon has also been tested systemically and topically against HSVbut with only limited efficacy. There are so many side effectsassociated with systemic administration of anti-virals that topicaltreatments are greatly preferred. Unfortunately, the topicaladministration even of the compounds having some efficacy whenadministered systemically is generally ineffective.

It is therefore an object of the present invention to provide topicalcompositions and methods of use thereof for the alleviation of thesymptoms associated with herpes simplex infections or other virallesions.

It is a further object of the present invention to provide topicalcompositions for alleviation of the symptoms of HSV infection withminimal side effects and relief from inflammation and pain normallyassociated with viral lesions.

SUMMARY OF THE INVENTION

The present invention provides a topical composition for reducing thesymptoms of viral lesions or shortening time of healing. The compositioncomprises a therapeutically effective amount of the combination of ananesthetic, a surfactant and a topical carrier. Varying amounts ofanesthetic and surfactant may be used to achieve efficacious results,e.g., for anesthetic concentrations of from about 0.05% to 25% byweight, 0.25% to 10% by weight, and most preferably 1 to 5% by weight,and for surfactant, concentrations of from about 0.05 to 50% by weight,1 to 10% by weight, and 0.5 to 5% by weight.

In preferred embodiments, the anesthetic is selected from the groupconsisting of esters, amides, ethers, and combinations thereof and, inparticular, topical anesthetics and other anesthetics which may beformulated in accordance with the invention and applied topically,including procaine, chloroprocaine, tetracaine, propoxycaine,benzocaine, cocaine, proparacaine, bupivacaine, dibucaine, etidocaine,lidocaine, mepivacaine, prilocaine, dyclonine, pramoxine andcombinations thereof.

The surfactant is selected from the group consisting of anionic,nonionic, and cationic surfactants and combinations thereof. Suitableionic surfactants include anionic surfactants such as monovalent salts,e.g., sodium and potassium salts of alkyl, aryl and alkyl-aryl sulfatesand sulfonates, particularly those with from about 8 to about 22 carbonatoms, and cationic surfactants, such as quaternary ammonium salts.Suitable nonionic surfactants include polyethylene oxide adducts offatty alcohols, e.g., alkylated polyoxyethylenes, alkylatedpolyoxyethylene-polyoxypropylene copolymers, and the surfactantnonoxynol.

The surfactant and the anesthetic are formulated together in apharmaceutically acceptable topical carrier, which may take any of anumber of acceptable forms. Suitable carriers include aqueous carriersand oleaginous carriers. The composition may contain one or moreadditional agents, including antimicrobial agents, anti-viral agents,anti-fungal agents, buffering agents, antioxidants, preservatives,coloring agents, fragrances, lubricants, moisturizers, sunscreens,drying agents and the like and, more specifically, may includeingredients such as stearic acid, lauramide DEA, borax, eucalyptus oil,beeswax, preservative and methylparaben.

Optimally, the composition will be effective to reduce the viral titerof lesions resulting from a viral disease and, in particular, herpeticlesions. The invention also includes methods for reducing the symptomsof viral lesions or decreasing time of healing by topicallyadministering compositions of the invention to a lesion or affectedarea, preferably at least about once every twelve hours and, morepreferably, about every four hours. The composition is applied to thelesions daily, preferably beginning at the preliminary or prodromalstage of infection. Relief is almost immediate, and is characterized bydecreased pain, swelling, and more rapid healing as compared with theuntreated lesion.

DESCRIPTION OF THE DRAWINGS

FIG. 1: FIG. 1 is a Kaplan-Meier analysis comparing the times to healingof heated versus control lesions.

FIG. 2: FIG. 2 is a plot the percentage of subjects who healed on adaily basis.

DETAILED DESCRIPTION OF THE INVENTION

The formulation for alleviation of the symptoms of HSV lesions has twoessential components, a topical or local anesthetic and a surfactant,which are added to a conventional carrier cream, ointment, solution orgel containing appropriate stabilizers, buffers and preservatives fortopical application to the lesion.

Topical anesthetics conveniently can be classified as esters, ethers oramides and include the following anesthetics which may be useful for thepractice of the invention:

    ______________________________________                                        Esters          Amides      Ethers                                            ______________________________________                                        Tetracaine      Dibucaine   Dyclonine                                         Benzocaine      Lidocaine   Pramoxine                                         Proparacaine                                                                  Procaine                                                                      Cocaine (not a legal drug in the United States)                               Butambin picrate (trinitrophenol 2-butyl-p-aminobenzoate)                     Propoxycaine                                                                  ______________________________________                                    

There are other anesthetics used for injection, e.g., spinal anesthesia,which are not usually listed as topicals but are expected to work wellon lesions where intact skin does not present a barrier.

Some examples are:

    ______________________________________                                        Chloroprocaine       Bupivacaine                                                                   Etidocaine                                                                    Mepivacaine                                                                   Prilocaine                                               ______________________________________                                    

The preferred anesthetic is tetracaine (either as the hydrochloride orother salt or as the free base). Tetracaine is the preferred anestheticbecause it is the anesthetic first tested and demonstrated to penetrateskin effectively, it has in vitro viral anti-replication activity, andit is on the FDA monograph list which allows marketing as anover-the-counter (OTC) product for fever blisters and cold sores atconcentrations up to 2% by weight.

In the preferred embodiment, the anesthetic is combined with a carrierin a concentration range of between 1 and 2% by weight; however, theanesthetic may be present in varying concentrations, for example,between approximately 0.05 and 25%, by weight, between approximately0.25 and 10% by weight, between approximately 0.5 and 10% by weight, andbetween approximately 1 and 5% by weight. Of course, the exact dose ofanesthetic to be employed in a given formulation will depend on a numberof factors such as the particular anesthetic to be employed. Forexample, in compositions containing benzocaine as the anesthetic, anexemplary preferred dose range is between approximately 1 and 20% byweight, in compositions containing lidocaine, an exemplary preferreddose range is between approximately 1 and 20% by weight and incompositions containing tetracaine an exemplary preferred dose isapproximately 0.5%.

In addition to the local anesthetic, the compositions of the inventionwill also contain a therapeutically effective amount of a surfactant.Surfactants have been discovered to have potent virucidal activity invitro with a very low degree of toxicity. It is believed that thesurfactant may both physically inactivate viruses and may facilitatepenetration of the anesthetic to the deeper, infected layers of theskin, enhancing the effectiveness of the individual compounds tofacilitate healing.

In a preferred embodiment, the surfactant in the formulation is sodiumlauryl (or dodecyl) sulfate, also referred to as "SLS", a commonsurfactant found in toothpaste and other cosmetic preparations, known to"unfold" the tertiary structure of proteins but not previously known tohave actual virucidal activity. Sodium lauryl sulfate is an anionicsurfactant with a negatively charged sulfate group. Sodium laurylsulfate is also capable of penetrating intact skin very effectively.Other monovalent sulfur-containing surfactants, e.g., the sodium orpotassium alkyl, alkyl-aryl, and aryl sulfates or sulfonates can be usedin accordance with the invention.

In addition, cationic surfactants may be active, alone or in combinationwith SLS. An example is trimethyldodecylammonium chloride, a positivelycharged quaternary ammonium complex which has antimicrobialcharacteristics. Other quaternary salts, with and without long chainmoieties to provide surface activity, may also be useful.

Nonionic surfactants, the most common being nonoxynol, may also beuseful. Common nonionic surfactants include nonoxynol, polyoxyethylenes,polyoxyethylene-polyoxypropylene copolymers and alkyl sorbitols.

In the preferred embodiment, the surfactant is combined with carrier ina concentration range of between approximately 0.5 to 2%, by weight;however, the surfactant may be present in a concentration of betweenapproximately 0.1 to 10%, by weight. The selected surfactants may alsobe combined in varying concentration ranges, for example, betweenapproximately 0.1 to 20% by weight, between approximately 0.05 to 50% byweight, between approximately 1.0 and 10% by weight, and betweenapproximately 0.5 and 5.0% by weight.

The composition can also contain antimicrobials, including antibiotics,antifungals, and other anti-viral compounds, which may complement orsupplement the activity of the basic composition. Suitable antibioticsinclude tetracycline, polymyxin B or other common antibiotics used intopical compositions, especially over-the-counter formulations. Examplesof useful antifungals include tolnaftate and micatin. Examples ofanti-virals include interferon, either natural or recombinant, as wellas nucleoside analogs, e.g., Acyclovir. Counter-irritants such ascamphor and menthol, drying agents such as benzyl alcohol, resorcinoland phenol, and astringents such as zinc sulfate and tannic acid canalso be added to the composition as can other types of agents such assunscreens, emollients, preservatives, fragrances, antioxidants, coloradditives, lubricants, moisturizers or drying agents. For example, asunscreen, e.g., PABA, can be added to the formula since it is knownthat cold sores can be triggered by ultraviolet radiation.

The composition can be prepared in almost any relatively inert topicalcarrier. Generally, the formulation could take several forms, e.g.,cream, gel, ointment, "Chapstick" and solution forms. Each of theseformulations may contain the two active ingredients as well asmicroorganism growth inhibitors (preservatives). Many such carriers areroutinely used and can be obtained by reference to pharmaceutical texts.Examples include polyethylene glycols (PEG), polypropylene copolymers(Pluronics), and some water soluble gels.

The preferred carrier is an emulsified cream, but other common carrierssuch as certain petrolatum or mineral oil-based ointments in which thesurfactant and anesthetic are dispersible can be substituted. Forexample, one suitable cream formulation used in animal studies andclinical experiments is described below:

    ______________________________________                                        COMPONENT      PERCENT (approximate)                                          ______________________________________                                        Deionized water                                                                              69                                                             Stearic Acid   22                                                             Sodium lauryl sulfate                                                                        1                                                              Beeswax        1                                                              Tetracaine     2                                                              Borax          0.4                                                            Lauramide DEA  3.6                                                            Methylparaben  0.3                                                            Eucalyptus Oil 0.03                                                           ______________________________________                                    

At this time, the preferred composition of the composition mosteffective in enhancing healing of viral lesions is: (all percents are byweight):

methylparaben, 0.25% (0.25 to 0.5%)

borax, 0.5% (0.3 to 0.6%)

lauramide DEA, 4% (0.3 to 0.4%)

stearic acid, 20% (15 to 25%)

beeswax, 0.8% (0.5 to 1.5%

tetracaine, 1.8% (1 to 2%)

sodium lauryl sulfate, 0.8% (0.1 to 3%)

eucalyptus oil 0.025% (0.01 to 0.0.5%)

PCMX 0.4% (0.3 to 0.5%)

water to a total of 100%

Gels, i.e., thickened aqueous or alcoholic solutions, containing theactive ingredients and stabilizers may be clear and/or colored withsuitable dyes. Suitable thickeners may include carboxymethylcellulose,polyvinylpyrrolidone or polyacrylic acid salts.

Hydrophilic or hydrophobic ointments may be employed as carriers.However, hydrophobic ointments, such as Vaseline, which are based uponhydrocarbon and wax derivatives may not be as efficacious as thehydrophilic ointments because they may impede penetration into the skin.Hydrophilic ointments such as those based upon propylene glycol,polyalkylene glycols, and the Pluronics are therefore preferred forointment formulations. Propylene glycol, as a base, is preferable topolyethylene glycol.

"Chapstick" formulations may be employed in situations, e.g., fortreatment of cold sores, where ease of application is a primaryobjective.

Solutions, i.e., dilute aqueous preparations containing activeingredients and preservatives but without substantial concentrations ofthickeners, can be sprayed upon the affected surface with an aerosolpump. This type of delivery may be of value for treating larger areas ofpainfully sensitive skin in indications such as shingles.

In most cases, it is preferred that the pH of the carrier containing theactive ingredients is adjusted to a pH of about 6 to 7, using, asbuffering agents, ingredients such as borax although other acceptablebuffering agents could be used.

Additional additives may include antioxidants, fragrance, color, water,preservatives (either antioxidants or antimicrobials), lubricants,moisturizers, or drying agents.

The composition is applied by the patient to the lesions one to sixtimes daily, most preferably beginning immediately after the prodrome issensed. Enough composition is applied to cover the lesion or theprodromal area.

The formulation has been demonstrated to be active against both HSV 1(cold sores and fever blisters) and HSV 2 (genital herpes). However, itmay be active against other viruses including herpes zoster (causingshingles) and the varicella virus causing chicken-pox, which are similarin structure or mechanism of infection.

The invention will be further understood by reference to the followingexamples:

EXAMPLE 1

Determination of the Ability of Selected Components of the Compositionto Directly Inactivate or Inhibit the Replication of Herpes SimplexVirus Type-2.

Cells

A continuous line of human embryo lung cells (MRC-5) were grown inEagle's minimal essential medium (MEM) in Earle's salts supplementedwith 2 mM L-glutamine, 10 mg/ml gentamicin, 0.25 mg/ml amphotericin B(Fungisone) and 10% heat-inactivated fetal calf serum (growth medium).Cells were grown at 37° C. in a humidified 5% CO₂ atmosphere or inclosed flasks. Maintenance medium was the same basic composition, exceptthe concentration of fetal calf serum was 2%. Vero cells (continuousline of green monkey kidney cells) were grown in the same manner asMRC-5 cells.

Viruses

A clinical isolate from a genital lesion of herpes simplex virus, Type 2(HSV-2), was used.

Stocks of viruses were grown by infecting MRC-5 cells at a multiplicityof 0.01 plaque forming units (PFU) per cell and harvested in maintenancemedium. Virus was harvested by three cycles of freezing at -70° C. andthawing, followed by centrifugation at 1000×g at 4° C. for 15 minutes toremove cell debris. Resulting virus stocks were aliquoted and frozen at-70° C.

Chemicals

Stock solutions of procaine hydrochloride, niacin, sodium laurylsulfate, and tetracaine were prepared from commercially availablereagents. Stock solutions were prepared in Eagle's MEM withoutsupplements and stored at 4° C.

Cytotoxicity Studies

Cytotoxicity assays were performed in MRC-5 cells and in Vero cells. Thetest compound was dissolved in maintenance medium at selectedconcentrations and inoculated onto MRC-5 or Vero cells in 96 well tissueculture plates. Each concentration was inoculated in triplicate. Theplates were incubated at 37° C. in 5% CO₂ and graded for cytotoxicity at24, 48 and 72 hours after inoculation. The highest concentration ofcompound showing 50% cytotoxicity after 72 hours was designated the endpoint.

These studies yielded the following results. Procaine HCl exhibitedsignificant cytotoxicity in MRC-5 cells at concentrations above 0.25%after 24 hr incubation. Procaine HCl was not as cytotoxic in Vero cellsas it was in MRC-5 cells, showing significant toxicity only atconcentrations above 2.0%. Tetracaine HCL was also more toxic to MRC-5cells, showing significant toxicity at concentrations above 0.03% at 24hr, but not in Vero cells. Niacin was non-toxic in MRC-5 cells up toconcentrations of 0.25%. Sodium lauryl sulfate was highly toxic to MRC-5cells at concentrations above 0.0025%.

Inactivation Studies

To determine if a test compound directly inactivates HSV-2, 0.5 ml ofserial dilutions of the compound to be tested was mixed with 0.5 ml ofstock HSV-2 (titer=5.0×10 PFU/ml) and incubated at room temperature for30 minutes. After 30 minutes, the compound-virus mixture was diluted10,000 fold in maintenance medium and virus recovery was determined byinoculating MRC-5 cells. The cells were graded for cytopathic effectafter 24 hr, 48 hr, and 72 hr.

These studies yielded the following results. Procaine HCl did notdirectly inactivate HSV-2 at the concentrations tested, up to 4%.Tetracaine HCl did not directly inactivate HSV-2 at the concentrationstested, up to 2%. Niacin did not directly inactivate HSV-2 at theconcentrations tested, up to 0.25%. Sodium lauryl sulfate directlyinactivated HSV-2 at concentrations ranging from 30% down to 0.02% andpartially inactivated the virus at concentrations down to 0.0025%. Noinactivation occurred at a concentration of 0.00032%.

Anti-viral Studies

Anti-viral activity, i.e., the ability of a compound to inhibitreplication of virus in cells, was measured by infecting confluentmonolayers of MRC-5 cells in 12×100 mm culture tubes at a multiplicityof infection (MOI) of 3.0 plaque forming units (PFU) per cell of HSV-2by removing the maintenance medium, washing the monolayers three timeswith phosphate buffered saline (PBS), and adding 0.1 ml of HSV-1suspended in PBS. The multiplicity of infection was calculated bycounting the number of viable cells in a duplicate tube and diluting thestock virus to produce a viral inoculum in a volume of 0.1 ml whichcontained three times the PFUs as the number of cells counted. After a60 minute absorption period, the inoculum was removed and 4 mls of testcompound at appropriate concentrations in maintenance medium ormaintenance medium (control) was added.

The tubes were incubated at 37° C. for an additional 23 hours. At 24hours post-infection, the medium was removed and the monolayers werewashed twice with PBS and frozen and thawed 3 times in 1 ml ofmaintenance medium. Virus yields were determined by serial end-pointtitrations in 96-well tissue culture plates containing MRC-5 cells. Allexperiments were tested in triplicate, 50% end-points were calculatedand titers were expressed in TCID 50 units. Anti-viral effect wasdetermined by subtracting the difference between the TCID 50s of thetest compound and from the TCID 50s of HSV-2 virus stock titrated in thesame experiment and calculated in percentages.

These studies yielded the following results. Concentrations of procaineHCl up to 2% were unable to inhibit the replication of HSV-2. TetracaineHCl partially inhibited 90% of the replication of HSV-2 at 0.25% and0.5%. It is likely that higher concentrations of tetracaine HCl wouldcompletely inhibit the replication of HSV-2. Niacin did not inhibit thereplication of HSV-2. Sodium lauryl sulfate did not inhibit viralreplication at concentrations of 0.0025% and 0.0013%.

In summary, the results show that SLS is the most active ingredient ofthose tested as an inactivator of HSV-2 at concentrations ranging downto 0.02%, and that tetracaine HCl has significant anti-viral activity ata concentration of 0.5%, although procaine HCl does not. At highconcentrations, sodium lauryl sulfate may also act by desquamation ofinfected cells in the lesions, causing their rapid removal and enhancedhealing as a result.

EXAMPLE 2

Evaluation of the In Vivo Efficacy of Selected Components of theComposition on a Primary Genital Herpes Simplex Virus Type 2 Infectionof Guinea Pigs

The anti-viral activity of several compounds in a primary genital HSV-2infection of guinea pigs was tested. The experiments wereplacebo-controlled and uninfected animals were treated with each of thepreparations to assess skin irritation. Groups treated with 5% Acyclovirwere included as positive controls.

Materials and Methods

Genital HSV-2 Infection of Guinea Pigs

The MS strain of HSV-2 was utilized to infect animals. Female Hartleystrain guinea pigs, Charles River Breeding Laboratories, Kingston, N.Y.,weighing 250 to 300 g were inoculated intravaginally with 1.4×10⁵ plaqueforming units of HSV-2 one hour after being swabbed for the removal ofvaginal secretions.

Treatment of Guinea Pigs

Groups of ten infected guinea pigs were treated topically, 0.1 mlintravaginally and 0.1 ml on external genital skin, three times daily,approximately every eight hours, for seven days, beginning either 6 hror 48 hr after HSV inoculation. Groups of three uninfected animals wereused to assess toxicity of each preparation.

Sample Collection and Virus Assays

To determine the effect of treatment on vaginal virus replication, swabsof vaginal secretions were obtained on days 1, 3, 5, 7 and 10 afterHSV-2 inoculation, placed in a tube containing 2.0 ml of media, vortexedand frozen at -70° C. until titrated for HSV-2. When all samples werecollected, they were thawed, diluted serially and HSV-2 titersdetermined using a microtiter CPE assay in rabbit kidney cells. Meanpeak virus titers and areas under the virus titer-day curves werecalculated and analyzed.

Scoring of External Genital Lesions

To determine the effect of therapy on the development and spread ofexternal genital lesions, lesion severity was scored on a 0-5+ scalethrough the primary infection (21 days). Mean peak lesion scores and thearea under the lesion score-day curves were calculated and analyzed.

Evaluation of Efficacy

The number of animals infected over the number inoculated, lesionscore-day areas and virus titer-day areas under the curve, peak lesionscores and peak virus titers between untreated and placebo-treated orplacebo-treated and drug-treated animals were compared using theMann-Whitney U rank sum test. A p-value of 0.05 or less was consideredsignificant.

Samples

1A base formulation without procaine witih SLS

2A base formulation without procaine or SLS

3A base formulation without SLS, with 1.8% procaine

3 base formulation with 1.3% SLS and 1.8% procaine

5 base formulation without procaine, with 1.6% tetracaine and SLS

6 base formulation without procaine, with 1.6% tetracaine, interferon(Schering Plough, 200,000 units) and SLS.

The base formulation is as defined for the preferred embodiment of thecomposition for enhancing heating of viral lesions, but without SLS oranesthetic. Acyclovir (ACV) was used as a positive control (i.e., havinganti-viral activity). Placebo (PBS) treated animals were compared withan untreated control animal. 2A was compared with the untreated animalsand placebo-treated animals. ACV groups were compared to theplacebo-treated group. All others were compared to 2A.

Results

For primary irritation determination, groups of three guinea pigs weretreated with each of the samples or controls. No visible signs of anyskin irritation or genital toxicity were observed. The animals remainedhealthy and normal in appearance throughout the study.

Effect on Viral Replication

After HSV-2 inoculation, viral replication in the vaginal tract reachesa peak on days 3 to 5, then declines gradually with most animals havingcleared the virus by day 10. Only 5% ACV given 6 hr after viralinoculation significantly reduced the virus titer-day area under thecurve (AUC) or the mean peak virus titer (p-values of 0.05 and <0.01,respectively). Evaluation of topical anti-viral therapy on vaginal virustiters is summarized in Table 1. All other formulations failed to altervaginal viral replication, regardless of when therapy was initiated.

Effect on Lesion Development

Three to four days after HSV-2 inoculation, vesicular lesions begin toappear on the external genital skin. Lesions progressed to an ulcerativestage by days 7 to 8 and gradually healed by days 15 to 21. Animalstreated with the PBS placebo at +6 hr had a significantly increasedlesion score-day AUC value when compared to the untreated control group(p-value of <0.05). The vehicle control for the formulations (2A) givenat +6 hr had a significantly lower lesion score-day AUC value whencompared to the untreated control group (p-value of <0.05). The vehiclecontrol for the formulations (2A) given at +6 hr had a significantlylower lesion score-day AUC when compared to the +6 hr placebo and theuntreated control groups (p-value of <0.05 and 0.001 respectively).However, 2A initiated 48 hr post-inoculation resulted in a significantlygreater lesion score-day AUC when compared to both the +48 hr placeboand untreated control groups (p-values of <0.01 and 0.001 respectively).

Therapy with 1A, 5, 6 or 5% ACV initiated 6 hr following HSV-2inoculation significantly decreased lesion development (p-values of<0.05 or less). ACV 5% also reduced the mean peak lesion score (p-valueof <0.001). Therapy with these formulations at +48 hr failed to inhibitlesion development. Increased lesion score-day AUC's and mean peakscores were seen with 1A and 5 (p-values of 0.06 or less).

It is important to note that the placebo showed a lesion score of 42.6.Samples 1A, 3, 5, and 6, all containing 1.3% SLS, had scores of 17.2,31.7, 19.8, and 17.4, respectively, while samples 2A and 3A, notcontaining SLS, had scores of 27.8 and 21.0, respectively. Samples 3 and3A had atypical scores, believed to be due to errors in formulation ortesting. However, it is evident that the presence of SLS in theformulation reduces the severity of the lesion course in this model.

                  TABLE 1                                                         ______________________________________                                        EVALUATION OF TOPICAL ANTI-VIRAL THERAPY ON                                   INFECTION RATES AND VAGINAL VIRUS TITERS DURING A                             PRIMARY GENITAL HSV-2 INFECTION OF GUINEA PIGS                                                    Virus                                                                         Titer-                                                                        Day            Mean                                                 # Virus   Area           Peak                                                 Positive/ Under          Virus                                      Treatment.sup.a                                                                         # Inoculated                                                                            Curve    p-Value                                                                             Titer p-Value                              ______________________________________                                        Control       10/10     23.4   --    4.2   --                                 Placebo                                                                               +6 h  10/10     23.9   NS.sup.b                                                                            4.4   NS                                 (PBS)                                                                         2A      +6 h  10/10     22.6   NS    3.7   NS                                 1A      +6 h  10/10     22.1   NS    3.6   NS                                 3A      +6 h  10/10     21.7   NS    3.6   NS                                 3       +6 h  10/10     24.0   NS    3.5   NS                                 5       +6 h  10/10     22.1   NS    3.6   NS                                 6       +6 h   9/10     20.5   NS    3.2   NS                                 ACV 5%  +6 h  10/10     14.8   0.05  2.9   <0.01                              Placebo                                                                              +48 h  10/10     23.0   NS    4.0   NS                                 (PBS)                                                                         2A     +48 h  10/10     25.8   NS    4.1   NS                                 1A     +48 h  10/10     26.3   NS    4.3   NS                                 3A     +48 h  10/10     26.3   NS    4.3   NS                                 3      +48 h  10/10     25.3   NS    4.1   NS                                 5      +48 h  10/10     27.3   NS    4.0   NS                                 6      +48 h  10/10     26.6   NS    4.3   NS                                 ACV 5% +48 h  10/10     16.3   NS    4.2   NS                                 ______________________________________                                         .sup.a. Treatment was initiated at the times indicated and continued thre     times daily for seven days both topically and intravaginally.                 .sup.b. NS = Not statistically significant when compared to the untreated     control or appropriate placebotreated group. The placebotreated animals       were compared to the untreated control. 2A was compared to the placebo an     the untreated control. ACV groups were compared to the placebotreated         group. All others were compared to 2A.                                   

                  TABLE 2                                                         ______________________________________                                        EVALUATION OF TOPICAL ANTI-VIRAL THERAPY                                      ON LESION DEVELOPMENT DURING A PRIMARY                                        GENITAL HSV-2 INFECTION OF GUINEA PIGS                                                  Lesion Score-                                                                 Day Area            Mean Peak                                       Treatment.sup.a                                                                         Under Curve                                                                              p-Value  Score   p-Value                                 ______________________________________                                        Control       35.8       --     3.1     --                                    Placebo                                                                               +6 h  42.6       <0.05↑                                                                         3.4     NS.sup.b                              (PBS)                                                                         2A      +6 h  27.8       <0.05.sup.c ↓                                                                 2.8     NS                                    1A      +6 h  17.2       <0.01↓                                                                        2.0     NS                                    3A      +6 h  21.0       NS     2.8     NS                                    3       +6 h  31.7       NS     3.2     NS                                    5       +6 h  19.8       <0.05↓                                                                        2.5     NS                                    6       +6 h  17.4       <0.01↓                                                                        2.3     NS                                    ACV 5%  +6 h  4.9        <0.001↓                                                                       0.8     <0.001↓                        Placebo                                                                              +48 h  39.2       NS     3.1     NS                                    (PBS)                                                                         2A     +48 h  51.2       <0.01.sup.c ↑                                                                  3.8     NS.sup.d                              1A     +48 h  59.9       <0.01↑                                                                         4.3     <0.05↑                          3A     +48 h  56.2       NS     4.0     NS                                    3      +48 h  46.8       NS     3.7     NS                                    5      +48 h  56.4       0.06↑                                                                          4.2     <0.05↑                          6      +48 h  49.7       NS     3.9     NS                                    ACV 5% +48 h  41.7       0.07↑                                                                          3.7     NS                                    ______________________________________                                         .sup.a. Treatment was initiated at the times indicated and continued thre     times daily for seven days both topically and intravaginally.                 .sup.b. NS = Not statistically significant when compared to the untreated     control or appropriate placebotreated group. The placebotreated animals       were compared to the untreated control. 2A was compared to the placebo an     the untreated control. ACV groups were compared to the placebotreated         group. All others were compared to 2A.                                        .sup.c. p value was also 0.001 when compared to the untreated control.        .sup.d. p value was also 0.05 when compared to the untreated control.    

EXAMPLE 3

Additional Evaluation of the In Vivo Efficacy of Selected Components ofthe Composition on a Primary Genital Herpes Simplex Virus Type 2Infection of Guinea Pigs

This example sets forth the results of additional studies to establishthe efficacy of the compounds in the guinea pig model.

The sodium lauryl sulfate (SLS) and tetracaine preparations tested werepurchased from Sigma Chemical Co., St. Louis, Mo. Preparations weredissolved and delivered in phosphate buffered saline (PBS) which wasalso the placebo vehicle control. The concentrations of each compoundutilized alone and in combination were 2%, 1%, and 0.2%.

Virus and Viral Inoculation

The MS strain of HSV-2 was utilized for the animal inoculation. FemaleHartley strain guinea pigs (Charles River Breeding Laboratories,Raleigh, N.C.) weighing 250-300 g were inoculated with 2.0×10⁵ plaqueforming units of HSV-2 one hours after being swabbed for the removal ofvaginal secretions.

Treatment of Guinea Pigs

Groups of 10 guinea pigs were treated both on the external genital skinand intravaginally (0.1 ml each route for a total of 0.2 ml pertreatment) for each preparation. Treatments were four times daily(approximately every 6 hours) for 14 days beginning 24 hours post-viralinoculation. Groups of 3 uninfected animals were treated as previouslystated with selected preparations to assess toxicity and skinirritation.

Sample Collection and Virus Assays

To determine the effect of treatment on vaginal viral replication, swabsof vaginal secretions were obtained on days 1, 3, 5, 7 and 10 after HSVinoculation, placed in a tube containing 2.0 ml of media and frozen at-70° C. until titrated for HSV-2. When all samples were collected, theywere thawed, vortexed, diluted serially and HSV-2 titers were determinedusing rabbit kidney cells in a microtiter CPE assay.

Evaluation of Efficacy

To determine the effect of therapy on the development and spread ofexternal genital lesions, lesion severity was scored on a 0-5+ scalethrough the primary infection (21 days). Lesion score-day areas andvirus titer-day areas under the curve, and peak lesion scores and peakvirus titers between untreated and placebo-treated or drug-treated andplacebo-treated animals were compared using the Mann-Whitney U rank sumtest. A p-value of 0.05 or less was considered significant.

Effect of Treatment with SLS, Tetracaine or Combinations of SLS plusTetracaine on Uninfected Guinea Pigs

No sign of local skin irritation from any of the formulations wasobserved. Throughout the treatment period, the genital skin remainednormal in appearance, and no redness or swelling was observed. Beginningon day 7 and for the duration of the treatment, there were periodicepisodes of an anesthesia-like effect observed in animals receiving 2%tetracaine, and some animals went to sleep. This effect was exhibitedapproximately 5 minutes after treatment was given and lasted about 20minutes. It was not always the same animals and not all animals wereaffected. Some animals went to sleep on several occasions, some times onthe same day and some times skipping several treatments. There seemed tobe no long term adverse affects following these incidents.

Effect of Treatment with SLS and/or Tetracaine on Vaginal Virus Titers

Viral replication in the vaginal tract after HSV-2 inoculation reaches apeak on days 3-5 and then declines with most animals having cleared thevirus by day 10. The effects of administration of the SLS and/ortetracaine preparations on vaginal virus titers are summarized in Table3. This experiment was designed to show whether or not viral titerreduction is enhanced by the combination of SLS and tetracaine comparedto the effects of the individual components. The combinations of SLS andtetracaine at 2% adn 1% levels provided significant reductions in viraltiter over the individual components at these levels. This effect wasnot apparent at the 0.2% concentration level. It is interesting to notethat SLS generally is more effective than tetracaine in reducing viraltiter compared to the placebo.

                  TABLE 3                                                         ______________________________________                                        EFFECT OF TREATMENT WITH SODIUM LAURYL SULFATE                                AND/OR TETRACAINE ON VAGINAL VIRUS TITERS OF                                  GUINEA PIGS INOCULATED INTRAVAGINALLY WITH HSV-2                                                   Virus                                                                         Titer-                                                                        Day           Mean                                                  # Virus   Area          Peak                                                  Positive/ Under         Virus                                      Treatment.sup.a                                                                          # Inoculated                                                                            Curve   p-Value                                                                             Titer p-Value                              ______________________________________                                        Placebo-PBS                                                                              10/10     28.4    --    4.6   --                                   SLS - 2%   10/10     19.6    NS.sup.b                                                                            3.9   <0.05                                SLS - 1%    7/10     18.5    0.05  3.1   <0.05                                SLS - 0.2%  7/10     13.8    <0.01 3.0   NS                                   Tetracaine - 2%                                                                           9/10     26.1    NS    4.0   NS                                   Tetracaine - 1%                                                                          10/10     29.6    NS    5.0   NS                                   Tetracaine - 0.2%                                                                         8/10     17.5    NS    3.6   NS                                   Tetracaine - 2% +                                                                         7/10     15.6    <0.05 3.0   NS                                   SLS - 2%                                                                      Tetracaine - 1% +                                                                         7/10     13.8    0.01  3.0   0.01                                 SLS - 1%                                                                      Tetracaine - 0.2% +                                                                       7/10     18.3    <0.05 3.0   NS                                   SLS - 0.2%                                                                    Tetracaine - 2% +                                                                         7/10     14.8    <0.05 3.2   NS                                   SLS - 1%                                                                      Tetracaine - 2% +                                                                        10/10     21.8    NS    3.9   NS                                   SLS - 1% cream                                                                ______________________________________                                         .sup.a. Topical and i.vag. treatment was initiated 24 hours after viral       inoculation and was continued four times daily for 14 days.                   .sup.b. NS = Not statistically significant when compared to the               placebotreated group.                                                    

Effect of Treatment with SLS, Tetracaine, or Combinations on LesionDevelopment

Four days after viral inoculation, vesicular lesions begin to appear onthe external genital skin. Lesions progress to an ulcerative stage bydays 7-8 and gradually heal by days 15-21. The effect of topicaltreatment with the SLS, tetracaine or combinations of the two on lesionseverity is shown in TABLE 4. Mean peak lesion scores were notsignificantly reduced for any preparation in this study. No otherdifferences on lesion development were observed in this study. Thisresult is explained by a deviation imparted to the standard protocol.Subjects were treated for fourteen rather than seven days. This extendedtreatment caused the lesions to remain moist (rather than drying outnormally) and so healing was impeded. The moist condition of the lesionsdid not affect the viral titer scores.

                  TABLE 4                                                         ______________________________________                                        EFFECT OF TREATMENT WITH SODIUM                                               LAURYL SULFATE AND/OR TETRACAINE                                              ON EXTERNAL LESION DEVELOPMENT IN A                                           GENITAL HSV-2 INFECTION OF GUINEA PIGS                                                   Lesion Score-                                                                 Day Area           Mean Peak                                       Treatment.sup.a                                                                          Under Curve                                                                              p-Value Lesion Score                                                                          pValue                                  ______________________________________                                        Placebo-PBS                                                                              24.1       --      2.5     --                                      SLS - 2%   23.3       NS.sup.b                                                                              2.5     NS                                      SLS - 1%   17.9       NS      2.7     NS                                      SLS - 0.2% 30.7       <0.05   2.7     NS                                      Tetracaine - 2%                                                                          18.5       NS      3.1     NS                                      Tetracaine - 1%                                                                          32.3       NS      3.3     NS                                      Tetracaine - 0.2%                                                                        15.5       0.01    1.5     NS                                      Tetracaine - 2% +                                                                        24.6       NS      2.8.5   NS                                      SLS - 2%                                                                      Tetracaine - 1% +                                                                        24.3       NS      2.6     NS                                      SLS - 1%                                                                      Tetracaine - 0.2% +                                                                      25.4       NS      2.4     NS                                      SLS - 0.2%                                                                    Tetracaine - 2% +                                                                        24.1       NS      2.8     NS                                      SLS - 1%                                                                      Tetracaine - 2% +                                                                        25.0       NS      2.9     NS                                      SLS - 1% cream                                                                ______________________________________                                         .sup.a. Topical and i.vag. treatment was initiated 24 hours postviral         inoculation and was continued four times daily for 14 days.                   .sup.b. NS = Not statistically significant when compared to the               placebotreated group.                                                    

EXAMPLE 4

Demonstration of In Vivo Efficacy in Enhancing Healing of HSV Lesions inHumans

Case report data for six patients was obtained in an open trial usingsamples containing SLS and procaine. It was shown that the averagenumber of days to healing was reduced to six from a historical mean often in the same patients. Furthermore, the course of the disease seemedto be altered favorably. The stages of infection are defined as follows:erythema (Ery); papule (Pap); vesicle (Ves); edema (Ede); and scab(Sca).

Table 5 provides the results of a clinical study in which the cream wasapplied topically four times daily to a recurring (not primary) lesion:

                                      TABLE 5                                     __________________________________________________________________________    Clinical Study Results                                                        Average                                                                       Duration                                                                      of Episode                            Episode                                 (Historical)                                                                              Infection                                                                           Number of Days      Duration                                Patient                                                                            Days   Type  Ery Pap Ves Ede Sca (days)                                  __________________________________________________________________________    1    10     genital                                                                             4   0   0   0   0   5                                       2    12     genital                                                                             5   2   2   2   1   6                                       3    10     labial                                                                              5   2   0   0   0   5                                       4    10     genital                                                                             4   3   0   0   0   4                                       5    14     genital                                                                             6   0   0   0   0   7                                       6     6     genital                                                                             8   0   2   4   3   8                                       Average                                                                            10.3 days                Average 5.8 days                                __________________________________________________________________________

It is important to note that four of the six patients never progressedbeyond the papule state and that the average days to healing was reducedby 44%, compared to historical controls which had been on systemicAcyclovir. Lesion-associated pain was reduced or eliminated in allcases.

EXAMPLE 5

Double Blinded Placebo Controlled Studies in Human Subjects SufferingFrom Herpes Labialis

In order to confirm the efficacy of the composition, additional largerscale studies were conducted in human patients. The studies weredesigned to assess the relative effectiveness and safety of compositionsof the invention versus placebo in the treatment of herpes labialis inimmunocompetent adults. The composition tested in these studies wasdesignated VT-33 and contained 1.8% tetracaine hydrochloride and 1%sodium lauryl sulfate.

The study was randomized, double-blind, and placebo controlled. Eligiblepatients were at least eighteen years of age and had prodromalsymptomatology or lesions of recurring herpes labialis. Patients inwhich lesions had been present for more than 48 hours (two days) priorto the initiation of study medication were not eligible for enrollment.Written informed patient consent was obtained.

After enrollment, the medication was self-administered every 2 wakinghours, until the lesions healed (scab loss), but for no more than 12days. A medical history, lesion examination, herpes simplex virusculture of the lesion, blood chemistries, complete blood count andurinalysis were performed on the day of enrollment prior to initiationof the study medication. Subjects were monitored for at least one visitduring their treatment and during a final visit after their lesions hadhealed.

Seventy-two patients were enrolled from four study centers. The patientsincluded in the study were male or female immunocompetent patients atleast eighteen years of age with a history of recurring herpes labialisand prodromal symptomatology or clinical signs of herpes labialis inwhich the lesion had been present for less than 48 hours prior to theinitiation of the study. Women of child bearing potential were enrolledprovided that they were not pregnant at entry and would not becomepregnant during the course of treatment.

The following patients were excluded from the study: (1) patients whocould not be evaluated until after lesions had healed, i.e., patientswho did not return for the final visit and complete theirself-evaluation forms; (2) those less than eighteen years of age; (3)those with a history of immediate hypersensitivity to tetracainehydrochloride; (4) pregnant or lactating females; (5) immunosuppressedindividuals, such as patients with immunodeficiency diseases (or thoseat high risk for developing immunodeficiency disease), malignanciesinvolving the immune system, or patients receiving immunosuppressanttherapy (including cyclosporin, AZT, 0KT-3, corticosteroids, alkylatingagents and or antimetabolites) within four months prior to presentationor radiation therapy within six months prior to presentation) orpatients likely to receive immunosuppressive or radiation therapy duringthe period of the study drug administration and or during the follow upperiod; (6) those who had been treated by any route (i.e., oral,topical, parenteral) with any anti-herpes or anti-viral agent within onemonth prior to or at the time of evaluation for enrollment; (7) thosewho had been treated with any investigational drug within one month ofevaluation for enrollment; (8) those with bacterial superinfection ofthe herpes simplex lesion at the time of evaluation for enrollment; (9)those whose herpes lesion had been present for more than forty eighthours (2 days) prior to initiation of study; (10) patients with aninability to tolerate the medication; and (11) patients with anycondition which, in the opinion of the investigator, would render thesubject unfit for the study.

Objective Evaluations

Several objective criteria were used to asses treatment efficacy.Results of these evaluations are set forth below.

Viral Cultures

Sixty-one of the 72 subjects (84.7%) tested positive for herpes simplexvirus by viral culture at enrollment.

Time to Scab Formation and Healing (Scab Loss)

Table 6 gives the results of the major objective evaluation parametersof the protocol used for the experiments: time to scab formation andtime to healing. There was no difference in the time to scab formationbetween the placebo and compound VT-33. However, there was astatistically significant difference in the healing time in subjectstreated with VT-33 and in subjects treated with the placebo. Subjectstreated with VT-33 healed on the average of 5.1 days, and those treatedwith placebo took longer to heal, on the average of 7.2 days (p=0.0002).This is a 29.2% reduction in healing time and indicates that VT-33 iseffective in treating recurrent herpes labialis.

                  TABLE 6                                                         ______________________________________                                        TIME TO SCAB FORMATION AND TIME TO HEAL OF                                    COMPOUND VT-33 VS. PLACEBO IN 72 SUBJECTS                                     (DAYS)                                                                                   PLACEBO  VT-33                                                                Mean ± SD (n)                                                                       Mean ± SD (n) p-Value                                  ______________________________________                                        TIME TO SCAB 2.4 ± 0.27 (34)                                                                       2.3 ± 0.26 (32) 0.4279                             FORMATION                                                                     TIME TO HEAL 7.2 ± 0.36 (33)                                                                       5.1 ± 0.35 (33) 0.0002                             ______________________________________                                    

FIG. 1 is a Kaplan-Meir analysis of the distribution of the times tohealing and graphically shows the difference between the placebo andVT-33 relative to the proportion of subjects still with scab over theperiod of treatment. Clearly, VT-33 treatment shifts the healing of thelesions to shorter times. Another graphic representation of this samedata is shown in FIG. 2, a plot of the percentage of subjects who healedon a daily basis. As seen the VT-33 curve is again shifted to shorterhealing times as compared to the placebo curve.

Subjective Evaluations

In addition to the objective observations shown above, subjectivepatient impressions were monitored during the study. Summary data onself-rating scales for pain relief by day on-study are shown in Table 7.Lower scores indicate less intense pain perceived by the subject. Therewere no statistically significant differences between treatment groupson self-ratings on pain intensity.

                  TABLE 7                                                         ______________________________________                                        Means, Standard Errors, and Sample Sizes of Individual                        Rating Scales for Relief of Pain by Day and Treatment Group.                               Placebo     Active                                               Day On-Study Mean ± S.E. (n)                                                                        Mean ± S.E. (n)                                   ______________________________________                                        1            3.8 ± 0.32 (32)                                                                        3.7 ± 0.33 (35)                                   2            3.2 ± 0.34 (33)                                                                        3.0 ± 0.28 (35)                                   3            2.8 ± 0.35 (33)                                                                        2.5 ± 0.29 (35)                                   4            2.2 ± 0.31 (33)                                                                        1.9 ± 0.21 (35)                                   5            1.8 ± 0.32 (32)                                                                        1.6 ± 0.19 (33)                                   6            1.8 ± 0.25 (31)                                                                        1.5 ± 0.14 (34)                                   7            1.5 ± 0.16 (29)                                                                        1.3 ± 0.10 (30)                                   ______________________________________                                    

Summary data on self-rating scales for intensity of itching by dayon-study are shown in Table 8. Lower scores indicate less intenseitching. There was statistically significantly lower self-rating ofitching reported by the Active group on days two and three of the study.

                  TABLE 8                                                         ______________________________________                                        Means, Standard Errors, and Sample Sizes of Individual                        Rating Scales for Relief of Itching by Day and Treatment Group.                            Placebo       Active                                             Day On-Study Mean ± S.E. (n)                                                                          Mean ± S.E. (n)                                 ______________________________________                                        1            3.1 ± 0.36 (32)                                                                          3.7 ± 0.47 (35)                                 2.sup.a,b    3.1 ± 0.42 (28)                                                                          2.3 ± 0.30 (34)                                 3.sup.a,b    2.7 ± 0.36 (31)                                                                          2.0 ± 0.26 (35)                                 4            2.0 ± 0.26 (29)                                                                          1.5 ± 0.16 (32)                                 5            1.6 ± 0.22 (29)                                                                          1.3 ± 0.12 (33)                                 6            1.4 ± 0.18 (29)                                                                          1.2 ± 0.07 (35)                                 7            1.3 ± 0.14 (25)                                                                          1.1 ± 0.06 (30)                                 ______________________________________                                         a-Student ttest, p < 0.05, onetailed.                                         bMann-Whitney Utest, p < 0.05, onetailed.                                

Summary data of self-rating scales for perceived benefit by day on-studyare shown in Table 9. Higher scores indicate a larger perceived benefit.There were no statistically significant differences between treatmentgroups on self-ratings of perceived benefit at the beginning of thestudy. However, as the study progressed the perceived benefit was ratedsignificantly higher by the Active group as compared to the Placebogroup on days three through seven.

                  TABLE 9                                                         ______________________________________                                        Means, Standard Errors, and Sample Sizes of Individual                        Rating Scales for Perceived Benefit by Day and Treatment Group.                            Placebo       Active                                             Day On-Study Mean ± S.E. (n)                                                                          Mean ± S.E. (n)                                 ______________________________________                                        1            3.1 ± 0.66 (18)                                                                          3.1 ± 0.60 (20)                                 2            4.5 ± 0.56 (31)                                                                          5.6 ± 0.47 (33)                                 3.sup.a,b    4.9 ± 0.55 (32)                                                                          6.5 ± 0.40 (31)                                 4.sup.a,b    5.4 ± 0.58 (31)                                                                          7.0 ± 0.44 (31)                                 5.sup.a      5.9 ± 0.59 (32)                                                                          7.1 ± 0.44 (31)                                 6.sup.a,b    5.4 ± 0.59 (32)                                                                          7.2 ± 0.45 (33)                                 7.sup.a,b    5.4 ± 0.61 (30)                                                                          7.6 ± 0.51 (29)                                 ______________________________________                                         a-Student ttest, p < 0.05, onetailed.                                         bMann-Whitney Utest, p < 0.05, onetailed.                                

Table 10 shows the subject's self-evaluation of the overall benefit oftreatment, where benefit was ranked on an index score of 1-10 (1=nobenefit; 10=very effective treatment) at the end of treatment. Subjectswho were treated with VT-33 ranked the overall benefit of theirtreatment at an index significantly higher than of the placebo-treatedgroup (7.3 vs. 5.9), which indicates that the VT-33 treated group had abetter subjective opinion of the medication than did the placebo-treatedsubjects.

                  TABLE 10                                                        ______________________________________                                        SUBJECT SELF-EVALUATION OF BENEFIT OF TREATMENT                               VT-33 VS. PLACEBO                                                             TREATMENT BENEFIT INDEX (n)                                                   ______________________________________                                        PLACEBO             5.9 ± .60 (34)                                         VT-33               7.3 ± .48 (37)                                         ______________________________________                                         p = .0359                                                                     INDEX                                                                         1 = No Benefit At All                                                         10 = Very Effective Treatment                                            

Investigator's Evaluation

The clinical investigators were also asked to evaluate the studymedications for their relative effects on the following subjectiveparameters: (1) relief from pain; (2) relief from itching; (3) abilityto heal; and (4) overall clinical effectiveness. Ranking was on an indexscale of 1-10 (1=very unfavorable; 10=very favorable). As shown in Table11, there was no statistical difference between the investigatorsranking in the above categories. One category, relief from itching,approached statistical significance.

                  TABLE 11                                                        ______________________________________                                        INVESTIGATOR'S EVALUATION OF RELIEF FROM PAIN,                                RELIEF FROM ITCHING, OVERALL HEALING EFFECT,                                  AND GENERAL CLINICAL BENEFIT (OF COMPOUND                                     VT-33 VERSUS PLACEBO)                                                                       PLACEBO (n) VT-33 (n)                                                                              p - VALUE                                  ______________________________________                                        RELIEF FROM PAIN                                                                            6.9 ± 0.55 (33)                                                                        7.8 ± 0.45                                                                          .0921                                      RELIEF FROM ITCHING                                                                         6.7 ± 0.57 (32)                                                                        7.9 ± 0.44                                                                          .0533                                      ABILITY TO HEAL                                                                             6.6 ± 0.63 (33)                                                                        6.9 ± 0.53                                                                          .3526                                      OVERALL CLINICAL                                                                            6.6 ± 0.60 (33)                                                                        7.3 ± 0.48                                                                          .1917                                      EFFECTIVENESS                                                                 ______________________________________                                         INDEX                                                                         1 = Very Unfavorable                                                          10 = Very Favorable                                                           No side effects or adverse reactions were reported in this study.        

No side effects or adverse reactions were reported in this study.

The results set forth above establish that compounds of the invention,as exemplified by compound VT-33, effectively shortened the healing timeof recurrent herpes labialis lesions by nearly 30% in a controlledclinical setting. Because of the design of this study, patients begantreatment as late as 48 hours after the appearance of their lesions.Therefore, it is possible that self-initiated therapy at the beginningof lesion appearance or at prodrome would result in a further shorteningof healing time. Additionally, the patients' impression of the benefitof the compound was significantly higher than that for the placebo. Themore conservative view of the investigators is contradicted by theobjective indications and patients' impressions, and therefore, may notrepresent an accurate assessment, particularly since the clinicians wereasked to evaluate parameters, such as pain and itching, that they didnot themselves experience.

Modifications and variations of the composition and method of thepresent invention will be obvious to those skilled in the art. Suchmodifications and variations are intended to come within the scope ofthe appended claims.

What is claimed is:
 1. A topical composition for reducing the symptomsof herpetic vital lesions or shortening time of healing of said lesionscomprising:(a) a therapeutically effective amount of the combinationof:(i) tetracaine present at a concentration of from about 0.25 % to 10%by weight; (ii) an anionic present at a concentration of from about 0.01to 50% by weight; and (b) a topical carrier.
 2. A topical compositionfor reducing the symptoms of herpetic viral lesions or shortening timeof healing of said lesions comprising:(a) a therapeutically effectiveamount of the combination of:(i) dibucaine present at a concentration offrom about 0.25% to 10% by weight; (ii) an anionic surfactant present ata concentration of from about 0.05% to 50% by weight; and (b) a topicalcarrier.
 3. A topical composition for reducing the symptoms of herpeticviral lesions or shortening time of healing of said lesionscomprising:(a) a therapeutically effective amount of the combinationof:(i) dyclonine present at a concentration of from about 0.25% to 10%by weight; (ii) An anionic present at a concentration of from about0.25% to 10% by weight; and (b) a topical carrier.
 4. A topicalcomposition for reducing the symptoms of herpetic viral lesions orshortening time of healing of said lesions comprising:(a) atherapeutically effective amount of the combination of:(i) bupivacainepresent at a concentration of from about 0.25% to 10% by weight; (ii) Ananionic surfactant present at a concentration of from about 0.05% to 50%by weight; and (b) a topical carrier.
 5. The topical composition of anyone of claims 1-2 and 3 wherein said topical carrier further comprisesone or more additional components selected from the group consisting of:stearic acid, lauramide DEA, borax, eucalyptus oil, beeswax,preservative, methylparaben and combinations thereof.
 6. The topicalcomposition of claim 5 wherein said composition is effective to reducethe viral titer of said lesions.
 7. The topical composition of any oneof claims 1-4 wherein said anionic surfactant is sodium lauryl sulfate.8. A topical composition for reducing the symptoms of herpetic virallesions or shortening time of healing of said lesions comprising:(a) atherapeutically effective amount of the combination of:(i) prilocainepresent at a concentration of from about 0.25% to 10% by weight; (ii) ananionic surfactant present at a concentration of from about 0.01 to 50%by weight; and (b) a topical carrier.